Objective To investigate the therapeutic effect of Xuebijing on anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis mice and its effect on the changes of helper T cell 1(Th1)/helper T cell 2(Th2).Methods The experimental groups included control group,model group,low-dose Xuebijing(low-XBJ)group,high-dose Xuebijing(high-XBJ)group,with 10 mice in each group,except the control group,the other 3 groups of mice were treated with antigen injection and immune stimulation to establish anti-NMDAR encephalitis models,the mice in the low-XBJ and high-XBJ groups were intraperitoneally injected with 5 ml/kg and 10 ml/kg of Xuebijing injection,respectively,once every 12 hours for 3 consecutive days;After 2 weeks,HE staining was used to observe the pathological changes of mice brain tissue,Nissl staining was used to observe the morphological changes of mice neurons,TUNEL staining was used to detect the apoptosis of mice neurons,ELISA was used to detect the contents of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-4 and interferon-γ(IFN-γ)in serum and cerebrospinal fluid of mice,immunohistochemical staining and Western blotting were used to detect the expression of IFN-γ and IL-4 in brain tissue,flow cytometry was used to detect the distribution of Th1 and Th2 cells in peripheral blood,and the ratio of Th1/Th2 was calculated.Results Compared with the model group,hippocampal tissue damage was improved in low-XBJ group and high-XBJ group,the morphological changes of neurons were small,the morphology of Nissl bodies was more complete and the number of Nissl bodies was increased,the TUNEL positive rate of neurons decreased,the contents of TNF-α,IL-1β and IFN-γ in serum and cerebrospinal fluid decreased while the content of IL-4 increased,the percentage of IFN-γ positive cells and the relative expression of protein in brain tissue decreased,the percentage of IL-4 positive cells and the relative expression of protein increased,the proportion of cluster of differentiation(CD)4+IFN-γ+labeled Th1 cells and the ratio of Th1/Th2 in peripheral blood decreased,and the proportion of CD4+IL-4+labeled Th2 cells in peripheral blood of mice in high-XBJ group increased,the differences were statistically significant(P＜0.05).In addition,compared with the low-XBJ group,the improvement effect of the above detection indicators in the high-XBJ group was more obvious,and the differences were statistically significant(P＜0.05).Conclusion Xuebijing can improve hippocampal neuronal damage in anti-NMDAR encephalitis mice,which may play a therapeutic role by reducing the expression of IFN-γ,promoting the expression of IL-4 and maintaining the balance of Th1/Th2 cells.

Objective To evaluate the effects of various polysorbates(PS)on the stability of different types of monoclonal antibody(mAb)drugs.Methods Three types of monoclonal antibodies mAbA(IgG1 proantibody drug),mAbB(IgG1 mAb)and mAbC(IgG1 mAb with Fc N297A mutation)were used as model proteins,and different kinds or contents of PS were added into the mAb formulations respectively to investigate the influencing factors.The effects of PS on the stability of mAb drugs were evaluated comprehensively by detecting the changes of quality attributes,such as protein aggregates and insoluble particles.Results PS20 and PS80 showed no significant difference in inhibiting the formation of aggregates and charge variants in the three mAbs(P>0.05),while the addition of PS80 in mAbB and PS20 in mAbC significantly inhibited the increase of insoluble particles respectively(P<0.05);The content of PS20 showed a significant effect on the detection indexes of charge variants and insoluble particles in mAbC(P<0.05).Conclusion Different types of mAbs have different sensitivities to various kinds and contents of PS.Therefore,when designing the formulation of mAbs,it is necessary to select appropriate kinds and contents of PS to further improve the stability of mAb drugs.

OBJECTIVE: To evaluate the associations of lipid indicators and mortality in Beijing Elderly Comprehensive Health Cohort Study. METHODS: A prospective cohort was conducted based on Beijing Elderly Comprehensive Health Cohort Study with 4499 community older adults. After the baseline survey, the last follow-up was March 31, 2021 with an average 8.13 years of follow-up. Cox proportional hazard model was used to estimate the hazard ratios (HR) with 95% CI for cardiovascular disease (CVD) death and all-cause death in associations with baseline lipid indicators. RESULTS: A total of 4499 participants were recruited, and the mean levels of uric acid, body mass index, systolic blood pressure, diastolic blood pressure, fasting plasma glucose, total cholesterol (TC), triglyceride, and low-density lipoprotein cholesterol (LDL-C) showed an upward trend with the increasing remnant cholesterol (RC) quarters (P_trend < 0.05), while the downward trend was found in high-density lipoprotein cholesterol (HDL-C). During the total 36,596 person-years follow-up, the CVD mortality and all-cause mortality during an average 8.13 years of follow-up was 3.87% (95% CI: 3.30%-4.43%) and 14.83% (95% CI: 13.79%-15.86%) with 174 CVD death participants and 667 all-cause death participants. After adjusting for confounders, the higher level of TC (HR = 0.854, 95% CI: 0.730-0.997), LDL-C (HR = 0.817, 95% CI: 0.680-0.982) and HDL-C (HR = 0.443, 95% CI: 0.271-0.724) were associated with lower risk of CVD death, and the higher level of HDL-C (HR = 0.637, 95% CI: 0.501-0.810) were associated with lower risk of all-cause death. The higher level of RC (HR = 1.276, 95% CI: 1.010-1.613) increase the risk of CVD death. Compared with the normal lipid group, TC ≥ 6.20 mmol/L group and LDL-C ≥ 4.10 mmol/L group were no longer associated with lower risk of CVD death, while RC ≥ 0.80 mmol/L group was still associated with higher risk of CVD death. In normal lipid group, the higher levels of TC, LDL-C and HDL-C were related with lower CVD death. CONCLUSIONS In community older adults, higher levels of TC and HDL-C were associated with lower CVD mortality in normal lipid reference range. Higher RC was associated with higher CVD mortality, which may be a better lipid indicator for estimating the CVD death risk in older adults.

Objective    To reveal the potential mechanism of cisplatin resistance in non-small cell lung cancer A549 cells by comparing the expression profiles of wild-type A549 cells and cisplatin-resistant A549 cells (A549/DPP) through whole transcriptome sequencing analysis. Methods    The cisplatin resistant A549 (A549/DDP) cell line was first established. Then, the whole-transcriptome analysis was conducted both on A549 and A549/DDP cells. Next, the differentially expressed RNAs of lncRNA-seq, circRNA-seq, and miRNA-seq data were identified, respectively, followed by functional enrichment analysis. Finally, a comprehensive analysis based on the whole transcriptome data was performed and the construction of the ceRNA network was carried out. Results    A total of 4 517 lncRNA, 123 circRNA, and 145 miRNA were differentially expressed in A549/DDP cells compared with the A549 cell line. These different RNAs were significantly enriched in cancer-related pathways. The ceRNA network contained 12 miRNAs, 4 circRNAs, 23 lncRNAs, and 9 mRNA nodes, of which hsa-miR-125a-5p and hsa-miR-125b-5p were important miRNAs based on the topological analysis. Conclusion    Tumor necrosis factor signaling pathway and p53 signaling pathway are involved in A549/DPP resistance. Hsa-miR-125a-5p and hsa-miR-125b-5p may be potential targets for reversing cisplatin resistance.

Objective To observe the enzymatic changes of myocardial sarcoplasmic reticulum in rats after injecting endotoxin (LPS), and provide basic research results for the future study of myocardial sarcoplasmic reticulum dysfunction caused by LPS in rats. Methods Ten SD rats were randomly divided into blank control group and LPS injection group with 5 rats in each group. In the LPS injection group, endotoxin was injected into the tail vessels of the rats. Results The heart rate (HR) of the LPS injection group increased and was faster than that in the blank control group [(204±18) beat per min vs. (139±10) beat per min on the first day, and (199±22) beat per min vs. (143±17) beat per min on the next day, both P values were less than 0.05]. The mean arterial pressure (MAP) was lower than that of the blank control group on the first day [(87±12) mmHg vs. (102±7) mmHg, P<0.05]. Under light and electron microscope, the myocardial cells of rats with LPS injection were loosely arranged, with intercellular infiltration with inflammatory cells, muscle fibers broken, and difficult to identify the morphology of mitochondria and sarcoplasmic reticulum. Quantitative PCR results showed that after endotoxin injection, troponin (CASQ1), sodium-calcium exchanger (NCX), calmodulin phosphatase 1 (ppplCa), phospholipid protein (PLN), sarcoplasmic reticulum Ca2+-ATPase (SERCA2) increased significantly (P<0.05). Conclusion Endotoxin can inhibit cardiomyocyte function by affecting the activity of sarcoplasmic reticulum calcium regulatory protein-related enzymes through various mechanisms.

Objective To characterize exosomes released from human umbilical vein endothelial cells (HUVECs) after heatstroke, and explore their effects on biological behaviors of cells. Methods HUVECs were cultured and divided into the control group and the heat shock group. The heat shock group cells were exposed at 41 ℃ for 2 hours; the control group cells were incubated at 37 ℃ for 2 hours. The characteristics of exosomes secreted by the two groups were analyzed by nanoparticle tracing. The expression of exosomes marker proteins, CD63 and TSG101, were detected by Western blotting. The process of exosomes entering normal cells was observed by fluorescence labeling. To further explore the potential function of exosomes secreted from the heat-shocked cells, cells were co-cultured with exosomes collected from cells cultured at 37 ℃ or cells cultured at 41 ℃. Cells under normal culture conditions were served as a negative control (blank control). FACS was used to detect cell cycle and apoptosis; MTS was used to evaluate cell proliferation; ELISA was used to quantify coagulation related factor expression; high-throughput sequencing was used to profile microRNAs (miRNAs) in the exosomes. Results Different from the control group, the cells in 41 ℃ heat shock group secreted abnormal exosomes, which resulted in abnormal behaviors and functions of normal HUVECs. The expression of tissue factor, von Willebrand factor, plasminogen activator inhibitor-1 increased significantly (P<0.05), and the expression of tissue plasminogen activator decreased (P<0.05). Compared with the incubation of HUVECs at 37 ℃, there were 2590 miRNA regulatory changes in exosomes secreted by HUVECs after heat shock at 41 ℃, which had obvious effects on apoptosis. Conclusions Heat shock at 41 ℃ could induce the release of abnormal exosomes from HUVEC, leading to abnormal behavior and potential coagulation of normal cells.

OBJECTIVETo investigate the effect of nucleolin silencing on the differentiation of rat neural stem cells (NSCs) and the role of Wnt signaling pathway in mediating such effect.

METHODSAdenovirus vectors expressing small interfering RNA (siRNA) against nucleolin were constructed, verified, and packaged in HEK293A cells. The adenovirus was then transfected into NSCs isolated from neonatal SD rats and the differentiation of the NSCs was examined by detecting the expressions of neuron specific encloase (NSE) and glial fibrillary acidic protein (GFAP) using immunocytochemistry. The expressions of nucleolin, nestin, Wnt3, and β-catenin in the cells were determined with Western blotting.

RESULTSRestriction endonuclease and sequencing analysis verified successful construction of the adenoviral vector expressing nucleolin siRNA (nucleolin-siRNA2). Infection of rat NSCs with nucleolin-siRNA2 significantly lowered nucleolin protein expression as compared with that in negative and blank control groups (P<0.05). The percentages of NSE-positive cells and GFAP-positive cells were significantly higher in NSCs infected with nucleolin-siRNA (P<0.01); the infection also resulted in obviously lowered expression of nestin protein and increased expressions of Wnt3 protein and β-catenin nucleoprotein in the cells.

CONCLUSIONSNucleolin silencing by adenovirus-mediated RNA interference induces the differentiation of NSCs into neurons and astrocytes, which is related with the activation of Wnt signaling pathway.

OBJECTIVETo compare the clinical effects of nasal intermittent positive pressure ventilation (NIPPV) and nasal continuous positive airway pressure (NCPAP) in the treatment of neonatal respiratory distress syndrome.

METHODSA prospective, randomized, controlled, single-center study was performed on 67 premature infants with NRDS between March 2011 and May 2012 and selected according to the inclusion and exclusion criteria. These premature infants were randomly assigned to receive NIPPV and NCPAP. Oxygenation index (OI), pH, PaCO2, duration of respiratory support, complications, success rate, hospital mortality, and incidence of bronchopulmonary dysplasia (BPD) were compared between the two groups.

RESULTSSixty-two patients were finally enrolled in the study, including 32 cases in the NIPPV group and 30 cases in the NCPAP group. After one hour of non-invasive ventilation, OI in the NIPPV group was higher than the NCPAP group (P<0.05), but there were no significant differences in pH and PaCO2 between the two groups (P>0.05 for both). A significantly lower proportion of infants needed mechanical ventilation via endotracheal tube (MVET) when they were treated initially with NIPPV than when they were treated initially with NCPAP (P<0.05). The NIPPV group had a significant higher success rate than the NCPAP group (P<0.05), but there was no significant difference in duration of respiratory support between the two groups (P>0.05). In addition, no significant differences in incidence of pneumothorax, hospital mortality and incidence of BPD were seen between the two groups (P>0.05 for all).

CONCLUSIONSCompared with NCPAP, NIPPV can significantly decrease the proportion of premature infants with NRDS in need of MVET. However, there is no evidence that NIPPV can significantly reduce hospital mortality and incidence of BPD in premature infants with NRDS.

Continuous Positive Airway Pressure ; Female ; Humans ; Infant, Newborn ; Intermittent Positive-Pressure Ventilation ; adverse effects ; Male ; Prognosis ; Prospective Studies ; Respiratory Distress Syndrome, Newborn ; therapy

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Heart Atria ; pathology ; Histiocytoma, Malignant Fibrous ; diagnosis ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo compare the compliance of using Actiwatch, parent report of sleep diary and sleep questionnaire in school age children, and to further evaluate agreement rates between actigraphy, diary, and questionnaire for children's sleep patterns.

METHODTwo primary schools in Luwan District were selected and first grade students without obvious physical and mental illnesses or sleep disorders were enrolled in the study. Each student was home-monitored with an Actiwatch for 7 days, meanwhile parents were asked to complete a detailed sleep diary during the Actiwatch monitoring days. Sleep questionnaires were distributed to their parents who were asked to fill in these on the last monitoring day.

RESULTForty-five children participated in the study, 36 children completed sleep assessments by all the three methods, and among them 20 were boys and 16 were girls. The mean age of those children was (7.26±0.42) years. The completion rate of questionnaire was 100%, of diary was 86.7% (39/45) and of Actiwatch was 93.3% (42/45). The completion rate between sleep diary and sleep questionnaire was significantly different (P<0.05). The satisfactory agreement between Actiwatch and sleep diary was reached with sleep diary overestimating weekday and weekend sleep duration by 26 minutes and 25 minutes respectively. The agreement rates between Actiwatch and questionnaire was insufficient for all variables with sleep questionnaire overestimating weekday and weekend sleep duration by 37 minutes and 38 minutes respectively.

CONCLUSIONSleep questionnaire is an easy and high compliance method for evaluating children's sleep pattern while sleep diary showed high agreement with Actiwatch. Using either of these methods for evaluating children's sleep pattern should be judged by research aim and sample size, and limitation of those methods should be considered when they are used in practice.

Actigraphy ; Child ; Female ; Humans ; Male ; Sleep ; physiology ; Surveys and Questionnaires